CD10+ neutrophils promote CD8+ T-cell depletion in mucosal malignant melanoma through the SELPG-SELL pathway

doi:10.3969/j.issn.1674-8115.2025.11.006

Abstract

Abstract:

Objective ·To compare the immune microenvironment of mucosal malignant melanoma (MM) and cutaneous malignant melanoma (CM) using single-cell transcriptome analysis system, and to elucidate the key regulatory mechanism of CD8⁺ T cell depletion in MM. Methods ·A total of 36 531 cells from three treatment-naïve MM surgical specimens and three CM samples were subjected to single-cell RNA sequencing. Data pre-processing included batch correction, strict quality control (based on mitochondrial gene ratio and gene number screening), unsupervised clustering, and cell-type annotation (using established marker genes). Differential gene analysis (Wilcoxon test), gene set enrichment analysis [based on Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases], cell-cell interaction network analysis, and prognostic association analysis using. The Cancer Genome Atlas (TCGA) were used to comprehensively analyse the heterogeneity of MM and CM tumour immune microenvironment. Results ·Compared with CM, the expression of cytotoxicity-related genes (GZMB, IFNG, etc.) in CD8⁺ T cells in MM was reduced (P<0.001), while the depletion markers (PDCD1, LAG3, etc.) were significantly up-regulated (P<0.001). The proportion of depleted CD8⁺ T cell subsets increased by nearly 6 times (CM: 3.38% vs MM: 19.26%, P<0.010), and the terminal depletion score was higher. In addition, the level of neutrophil infiltration in the MM microenvironment was significantly higher than that in CM (CM: 1.6% vs MM: 34.1%, P=0.009), and single-cell analysis further divided it into five subgroups. Among them, the CD10⁺ neutrophil subset (accounting for 23.52% of MM neutrophils) highly expressed pro-inflammatory molecules (S100A8/A9) and immunosuppressive molecules (MME/CD10, CD55), and the characteristic scores of myeloid-derived suppressor cells (MDSCs) in this subset were significantly higher than those of other subgroups. TCGA and MM cohort analyses further confirmed that the highly expressed genes of this subset were independently associated with poor prognosis (P=0.049), suggesting that this subset was involved in tumour immunosuppressive effects. Mechanistically, CD10⁺ neutrophils interacted with depleted CD8⁺ T cells through the SELPLG-SELL ligand-receptor pair, driving T-cell inactivation. Conclusion ·Single-cell transcriptome sequencing was performed on MM clinical samples, and the single-cell immunomes of MM and CM were systematically compared, suggesting that CD10⁺ neutrophils in MM induce CD8⁺ T-cell terminal depletion through the SELPLG-SELL pathway. The molecular mechanism of MM immunotherapy tolerance was elucidated, providing a new strategy for targeting neutrophil-T cell interaction.

Key words: mucosal malignant melanoma, neutrophils, CD8⁺depleted T cells, single-cell transcriptome

CLC Number:

R730.261

HAO Meiling, MA Yanni, MA Xuhui, ZHANG Yanjie, ZENG Hanlin, CHEN Shanshuang. CD10⁺ neutrophils promote CD8⁺ T-cell depletion in mucosal malignant melanoma through the SELPG-SELL pathway[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2025, 45(11): 1466-1479.

Figures/Tables 6

Fig 1 Single-cell mapping of mucosal malignant melanoma and cutaneous malignant melanoma

Fig 2 Comparison of T cell characteristics in the immune microenvironment of mucosal malignant melanoma and cutaneous malignant melanoma

Fig 3 Differences in the characteristics of neutrophil subpopulations in the microenvironment of mucosal and cutaneous malignant melanoma

Fig 4 Prognostic analysis of TAN2_CD10+ neutrophil subsets in the microenvironment of mucosal and cutaneous malignant melanoma

Fig 5 Characterization of T cell-neutrophil interaction networks in the microenvironment of mucosal and cutaneous malignant melanoma

Fig 6 TAN2_CD10+ neutrophil subpopulation is enriched by inflammation in the microenvironment of mucosal and cutaneous malignant melanoma

TrendMD

References 41

[1]	ZHOU L, CUI C L, LIAN B, et al. Postoperative radiotherapy in resected sinonasal mucosal melanoma[J]. J Clin Oncol, 2019, 37(15_suppl): e21059.
[2]	SALEH M, JAVADI S, ELSHERIF S, et al. Multimodality imaging and genetics of primary mucosal melanomas and response to treatment[J]. Radiographics, 2021, 41(7): 1954-1972.
[3]	OTTAVIANO M, GIUNTA E F, MARANDINO L, et al. Anorectal and genital mucosal melanoma: diagnostic challenges, current knowledge and therapeutic opportunities of rare melanomas[J]. Biomedicines, 2022, 10(1): 150.
[4]	KOTTSCHADE L A, POND G R, OLSZANSKI A J, et al. SALVO: single-arm trial of ipilimumab and nivolumab as adjuvant therapy for resected mucosal melanoma[J]. Clin Cancer Res, 2023, 29(12): 2220-2225.
[5]	SI L, FANG M Y, CHEN Y, et al. Atezolizumab in combination with bevacizumab in patients with unresectable locally advanced or metastatic mucosal melanoma: interim analysis of an open-label phase II trial[J]. J Clin Oncol, 2021, 39(15_suppl): 9511.
[6]	JOHNSON D B, SWETTER S M, SALAMA A K S, et al. Cutaneous melanoma: management of melanoma brain metastases and molecular testing[J]. J Natl Compr Cancer Netw, 19(5.5): 644-647.
[7]	LI S M, WU X W, YAN X Q, et al. Toripalimab plus axitinib in patients with metastatic mucosal melanoma: 3-year survival update and biomarker analysis[J]. J Immunother Cancer, 2022, 10(2): e004036.
[8]	BLACKBURN S D, SHIN H, FREEMAN G J, et al. Selective expansion of a subset of exhausted CD8 T cells by alphaPD-L1 blockade[J]. Proc Natl Acad Sci USA, 2008, 105(39): 15016-15021.
[9]	ZENG S S, HU H L, LI Z Y, et al. Local TSH/TSHR signaling promotes CD8⁺ T cell exhaustion and immune evasion in colorectal carcinoma[J]. Cancer Commun (Lond), 2024, 44(11): 1287-1310.
[10]	GAO Y, OUYANG Z J, YANG C, et al. Overcoming T cell exhaustion via immune checkpoint modulation with a dendrimer-based hybrid nano complex[J]. Adv Healthc Mater, 2021, 10(19): e2100833.
[11]	BELK J A, DANIEL B, SATPATHY A T. Epigenetic regulation of T cell exhaustion[J]. Nat Immunol, 2022, 23(6): 848-860.
[12]	KANG K, LIN X, CHEN P, et al. T cell exhaustion in human cancers[J]. Biochim Biophys Acta Rev Cancer, 2024, 1879(5): 189162.
[13]	MILLER B C, SEN D R, AL ABOSY R, et al. Subsets of exhausted CD8⁺ T cells differentially mediate tumor control and respond to checkpoint blockade[J]. Nat Immunol, 2019, 20(3): 326-336.
[14]	KALTENMEIER C, YAZDANI H O, MORDER K, et al. Neutrophil extracellular traps promote T cell exhaustion in the tumor microenvironment[J]. Front Immunol, 2021, 12: 785222.
[15]	GIESE M A, HIND L E, HUTTENLOCHER A. Neutrophil plasticity in the tumor microenvironment[J]. Blood, 2019, 133(20): 2159-2167.
[16]	WANG L W, LIU Y H, DAI Y T, et al. Single-cell RNA-seq analysis reveals BHLHE40-driven pro-tumour neutrophils with hyperactivated glycolysis in pancreatic tumour microenvironment[J]. Gut, 2023, 72(5): 958-971.
[17]	MENG Y, YE F, NIE P P, et al. Immunosuppressive CD10⁺ALPL⁺ neutrophils promote resistance to anti-PD-1 therapy in HCC by mediating irreversible exhaustion of T cells[J]. J Hepatol, 2023, 79(6): 1435-1449.
[18]	HUANG C T, LIAO Y H, LIAU J Y, et al. Pretreatment neutrophil-to-lymphocyte ratio predicts survival in primary mucosal melanoma[J]. J Am Acad Dermatol, 2025, 92(2): 340-343.
[19]	SADE-FELDMAN M, YIZHAK K, BJORGAARD S L, et al. Defining T cell states associated with response to checkpoint immunotherapy in melanoma[J]. Cell, 2018, 175(4): 998-1013.e20.
[20]	TIROSH I, IZAR B, PRAKADAN S M, et al. Dissecting the multicellular ecosystem of metastatic melanoma by single-cell RNA-seq[J]. Science, 2016, 352(6282): 189-196.
[21]	ZHANG C, SHEN H R, YANG T L, et al. A single-cell analysis reveals tumor heterogeneity and immune environment of acral melanoma[J]. Nat Commun, 2022, 13(1): 7250.
[22]	ZHOU Z Y, ZHOU X X, JIANG X, et al. Single-cell profiling identifies IL1B^hi macrophages associated with inflammation in PD-1 inhibitor-induced inflammatory arthritis[J]. Nat Commun, 2024, 15(1): 2107.
[23]	ALSHETAIWI H, PERVOLARAKIS N, MCINTYRE L L, et al. Defining the emergence of myeloid-derived suppressor cells in breast cancer using single-cell transcriptomics[J]. Sci Immunol, 2020, 5(44): eaay6017.
[24]	KELLIHER M A, RODERICK J E. NOTCH signaling in T-cell-mediated anti-tumor immunity and T-cell-based immunotherapies[J]. Front Immunol, 2018, 9: 1718.
[25]	LIU W, STACHURA P, XU H C, et al. BAFF attenuates immunosuppressive monocytes in the melanoma tumor microenvironment[J]. Cancer Res, 2022, 82(2): 264-277.
[26]	LIU X B, WANG Y Y, BAUER A T, et al. Neutrophils activated by membrane attack complexes increase the permeability of melanoma blood vessels[J]. Proc Natl Acad Sci USA, 2022, 119(33): e2122716119.
[27]	BARON M, IDEKER T. Abstract 178: desmosome mutations in melanoma promote cellular proliferation and disease progression[J]. Cancer Res, 2021, 81(13_Supplement): 178.
[28]	IRIONDO O, YU M. Unexpected friendship: neutrophils help tumor cells en route to metastasis[J]. Dev Cell, 2019, 49(3): 308-310.
[29]	GUNGABEESOON J, GORT-FREITAS N A, KISS M, et al. A neutrophil response linked to tumor control in immunotherapy[J]. Cell, 2023, 186(7): 1448-1464.e20.
[30]	WU Y C, MA J Q, YANG X P, et al. Neutrophil profiling illuminates anti-tumor antigen-presenting potency[J]. Cell, 2024, 187(6): 1422-1439.e24.
[31]	LIU S J, YUAN S J, LIU M C, et al. In situ tumor cell engineering reverses immune escape to enhance immunotherapy effect[J]. Acta Pharm Sin B, 2025, 15(1): 627-641.
[32]	BRANDAU S, HARTL D. Lost in neutrophil heterogeneity? CD10![J]. Blood, 2017, 129(10): 1240-1241.
[33]	SÁNCHEZ-MAGRANER L, MILES J, BAKER C L, et al. High PD-1/PD-L1 checkpoint interaction infers tumor selection and therapeutic sensitivity to anti-PD-1/PD-L1 treatment[J]. Cancer Res, 2020, 80(19): 4244-4257.
[34]	MA H Y, YAMAMOTO G, XU J, et al. IL-17 signaling in steatotic hepatocytes and macrophages promotes hepatocellular carcinoma in alcohol-related liver disease[J]. J Hepatol, 2020, 72(5): 946-959.
[35]	HAO Y W, LIU D L, WANG K Y, et al. Imaging and therapy of tumors based on neutrophil extracellular traps[J]. Small Sci, 2024, 4(10): 2400212.
[36]	YANG L Y, LUO Q, LU L, et al. Increased neutrophil extracellular traps promote metastasis potential of hepatocellular carcinoma via provoking tumorous inflammatory response[J]. J Hematol Oncol, 2020, 13(1): 3.
[37]	VIRAMONTES K M, NEUBERT E N, DEROGATIS J M, et al. PD-1 immune checkpoint blockade and PSGL-1 inhibition synergize to reinvigorate exhausted T cells[J]. Front Immunol, 2022, 13: 869768.
[38]	van NOT O J, van DEN EERTWEGH A J M, JALVING H, et al. Long-term survival in patients with advanced melanoma[J]. JAMA Netw Open, 2024, 7(8): e2426641.
[39]	de MEZA M M, van NOT O J, BLOKX W, et al. Efficacy of checkpoint inhibition in advanced acral melanoma[J]. J Clin Oncol, 2021, 39(15_suppl): e21527.
[40]	ZENG W Y, WANG Y, ZHANG Q, et al. Neutrophil nanodecoys inhibit tumor metastasis by blocking the interaction between tumor cells and neutrophils[J]. ACS Nano, 2024, 18(10): 7363-7378.
[41]	QUE H Y, FU Q M, LAN T X, et al. Tumor-associated neutrophils and neutrophil-targeted cancer therapies[J]. Biochim Biophys Acta Rev Cancer, 2022, 1877(5): 188762.